Lime peels, often discarded as waste in the food and beverages industry, encompass a vast range of indispensable bioactive compounds showing various therapeutic properties (e.g., antioxidant, bacteriostatic, antifungal, cytotoxic activities, etc.). This study aimed to optimize the drying conditions of Citrus aurantifolia peels dried using traditional (sun drying) and alternative drying methods (tray drying and vacuum drying), extract essential oil using hydro-distillation, and enhance its stability through nano-encapsulation with polyethylene glycol (PEG) 4000. Tray drying at 50 °C preserved the highest levels of phenolics (73.41 ± 0.15 mg GAE/g), flavonoids (45.52 ± 0.21 mg QE/g), radical activity (84.91 ± 0.09

Background:The changing epidemiology of candidemia indicates a rise in non-albicans Candida species, especially resistant Candida auris and emerging Candida utilis. Although iron impacts fungal virulence, its role in these species remains poorly understood. This study investigates how manipulating iron levels influences biofilm formation, virulence enzymes, and antifungal susceptibility in clinical isolates. Methods:A total of 216 isolates of Candida utilis, Candida albicans, and Candida auris from bloodstream infections over two years were identified via phenotypic methods, MALDI-TOF MS, VITEK 2, and 18S rRNA PCR. Susceptibility was tested using disc diffusion and broth microdilution with ferrous sulphate (FeSO4). Virulence enzyme activities and biofilm formation were assessed under iron-rich and control conditions. Results:Candida auris showed multidrug resistance, especially to fluconazole and caspofungin, with iron increasing caspofungin MICs up to 16-fold. Candida utilis exhibited strong biofilm formation and increased phospholipase and proteinase activities in the presence of FeSO4, and also showed 4- to 32-fold increases in fluconazole resistance. Biofilm biomass was unaffected by iron, but enzyme activities varied by species and enzyme. Candida albicans had high proteinase and haemolysin activity but responded minimally to iron. Conclusions:Iron differentially influences virulence-associated traits (biofilm-related enzyme activities) and antifungal resistance across these Candida species. C. utilis exhibits iron-responsive increases in phospholipase and proteinase activities together with amplified azole resistance, while C. auris shows iron-linked enhancement of echinocandin resistance and sustained expression of key virulence-associated enzymes. These results underscore the importance of accounting for host iron levels and species-specific responses when managing candidemia and indicate the potential for therapies targeting iron.

Aquaculture has become one of the fastest-growing food sectors globally, but its expansion is increasingly challenged by bacterial disease outbreaks and the rising threat of antimicrobial resistance (AMR). As conventional antibiotic treatments lose effectiveness, bacteriophage therapy – the use of viruses that selectively infect and lyse bacteria – has emerged as a promising alternative. Phages offer high specificity, minimal environmental impact, and proven efficacy against multidrug-resistant pathogens. This review provides an updated synthesis of current knowledge on the application of bacteriophages and phage-derived products, such as lysins and tail-like bacteriocins, in aquaculture systems. We explore their mechanisms of action, therapeutic advantages, and outcomes from experimental and field trials against key pathogens, including Aeromonas hydrophila, Vibrio harveyi, Edwardsiella tarda, and Streptococcus iniae. Importantly, phage therapy aligns with the One Health framework, which emphasizes the interdependence of human, animal, and environmental health. By reducing the use of antibiotics in aquatic farming, phage applications can help curb the spread of AMR, protect water ecosystems, and enhance food safety. Despite its potential, challenges such as phage resistance, endotoxin release, intracellular pathogen targeting, and standardization gaps must be addressed for broader adoption. We conclude by outlining future research priorities, including genome-guided phage selection, optimized delivery systems, and the need for standardized efficacy testing. Phage therapy thus represents a sustainable and integrative approach to aquatic disease management and global public health.

INTRODUCTION:Pseudomonas aeruginosa biofilms drive chronic infections, yet heterogeneous matrix composition hinders standardized in vitro classification of clinical isolates. This study aimed to develop a reproducible biofilm formation protocol using a defined supplement mix and elucidate proteomic signatures across biofilm categories to enable risk stratification and targeted therapies. METHODS:139 clinical P. aeruginosa isolates were tested in BHI broth supplemented with 0.5% each glucose, mannose, NaCl, and arginine. Biofilm formation was quantified via TCP assay (ODcut=0.344 at 570 nm), validated by confocal (ConA-TRITC), SEM, protein/eDNA quantification (Bradford, phenol-chloroform), SDS-PAGE, and LC-Orbitrap HRMS peptidomics (549 proteins; PRIDE PXD057726). Antimicrobial susceptibility followed CLSI guidelines. RESULTS:Supplement mix increased biofilm OD 26.9% (1.28±0.12 vs. 0.95±0.13; P<0.001), reclassifying isolates: HBF 38.8% (n=54), MBF 46.0% (n=64), WBF 13.7% (n=19), NBF 1.4% (n=2), resistance: ceftazidime 43.9%, meropenem 43.2%, imipenem 33.1%; 12.9% MDR. HBF matrices showed the highest protein levels (1.38±0.37 mg/mL); peptidomics revealed 12 shared proteins, 26 in PAO1/HBF/MBF, and 16 HBF-unique (ribosomal RpsA-RpsQ/RplA-RplY; stress YidC/KatA/ClpB/DnaJ; metabolic Edd/Pgk/PckA/ArcA). DISCUSSION:Distinct signatures indicate anoxic adaptations (HupB, AtpA-G) that drive HBF robustness, linking matrix heterogeneity(GeNei, India) to infection persistence beyond mere polysaccharide dominance. HBF matrices exhibited anoxic adaptation (HupB, AtpA-G), with YidC insertase facilitating membrane protein biogenesis under stress, supporting metabolic speciation for biofilm-specific diagnostics, therapies, and isolate risk stratification. CONCLUSION:This protocol enables biofilm categorization and reveals metabolic speciation targets for diagnostics and therapies against MDR P. aeruginosa biofilms.

Abstract Artificial intelligence (AI) is rapidly reshaping diagnostic microbiology by enhancing the speed, precision, and scalability of pathogen detection and antimicrobial susceptibility testing. Traditional approaches – such as manual microscopy, culture-based techniques, and biochemical assays – remain foundational but are often limited by long turnaround times, labor intensity, and subjective interpretation. In contrast, AI offers data-driven solutions capable of analyzing complex biological information, automating routine laboratory tasks, and improving clinical decision-making. This review explores the expanding applications of AI across multiple domains of diagnostic microbiology. We discuss the use of deep learning – particularly convolutional neural networks – for interpreting microscopy images, including Gram stain classification and parasite detection. We also highlight how machine learning models are transforming genomic diagnostics by predicting resistance phenotypes directly from whole genome sequencing and metagenomic datasets. Furthermore, the integration of AI into laboratory information systems is enabling workflow optimization, sample prioritization, and real-time support for antimicrobial stewardship through dynamic dashboards and alerts. Beyond current applications, AI is also opening new frontiers in outbreak forecasting, virulence prediction, and modeling of host–pathogen interactions using multiomics data. However, the integration of AI into clinical practice is not without challenges. Key concerns include data privacy, algorithmic bias, limited transparency of complex models, and evolving regulatory frameworks. Ultimately, AI should be viewed not as a replacement for microbiologists but as a powerful tool that augments human expertise. With responsible implementation and interdisciplinary collaboration, AI has the potential to transform microbiological diagnostics and advance global health outcomes.

ABSTRACT Aspergillus nidulans is a fungal pathogen that causes respiratory issues in individuals with compromised immune systems. It is identified through culture characteristics and microscopic features such as Cleistothecia and Hülle Cells. However, similar traits are found in cryptic Aspergillus species, such as stellatus, cristatus, and oryzae. In a case series from a hospital in North India, A. nidulans strains isolated from patients with pulmonary aspergillosis were phenotypically identified and then underwent molecular characterization through sequencing of the amplified ITS1‐5.8S rDNA‐ITS2 region and antifungal susceptibility testing (AST) according to the CLSI M38A3 guidelines. The molecular characteristics and antifungal profiles of the nine phenotypic A. nidulans were as follows: A. stellatus (n = 5), A. nidulans (n = 2), A. cristatus (n = 1), and A. oryzae (n = 1). Phylogenetic analysis revealed a close relationship between A. stellatus, A. nidulans, and A. cristatus, while A. oryzae showed significant divergence. Furthermore, the minimum inhibitory concentration (MIC) of antifungals was the lowest for caspofungin, followed by voriconazole. However, a higher amphotericin B MIC (2 µg/mL) was observed for A. stellatus. Thus, sequencing the ITS1‐5.8S rDNA‐ITS2 region can accurately identify cryptic species with superior taxonomic resolution. Additionally, the MIC of amphotericin B against A. stellatus underscores the importance of precise molecular identification and antifungal MIC profiling in each case of pulmonary aspergillosis.

Background: Endophytic fungi are known for diverse bioactive compounds with immense potential for agriculture and medicinal applications. Coniochaeta dendrobiicola isolated from the roots of Dedrobium longicornu was investigated for its antioxidant and metabolite composition. The present study compares the antioxidant properties, flavonoid and phenolic contents and metabolic profiles of broth and mycelium extracts. The broth and mycelium extracts were tested for their antioxidant potential using DPPH, while the total flavonoid and phenolic contents were measured using a UV–VIS spectrophotometer. High-resolution mass spectrometry (HRMS) revealed a markedly richer and more diverse metabolite profile of putatively annotated compounds in the broth extract compared with the mycelium fraction. The broth extract exhibited significantly higher antioxidant activity and flavonoid and phenolic contents, correlating with the presence of diverse bioactive compounds, including indole derivatives, flavonoids, phenolic acids, quinoline derivatives, and antifungal metabolites. Notably, several indole-related and phenolic compounds detected predominantly in the broth are known for antioxidant, antimicrobial, and plant growth-promoting properties. These findings indicate that C. dendrobiicola actively secretes biologically relevant secondary metabolites into the extracellular medium, highlighting its potential for agricultural and pharmaceutical applications.

A guar gum (GG)-grafted-(polydimethylamino-co-polyacrylamido sulfonic acid) [GG-g-(PDMAEA-co-PAMPS)] hydrogel was developed as a promising material for wound dressings. The hydrogel was synthesized by grafting poly(dimethylaminoethacrylate) (PDMAEA) and poly(acrylamidopropyl sulfonic acid) (PAMPS) onto guar gum (GG), and its structure was confirmed by Fourier transform infrared (FTIR) and X-ray diffraction (XRD) analyses. Rheological assessments demonstrated its mechanical robustness and self-healing properties while swelling studies revealed pH-sensitive behavior. Biocompatibility was confirmed through cell viability assays, showing minimal cytotoxicity and the hydrogel exhibited a bacteriostatic effect against Escherichia coli, Staphylococcus aureus, and Enterococcus faecalis. In a rat full-thickness chronic wound model, the hydrogel significantly accelerated wound healing, enhanced collagen deposition, reduced inflammation, and promoted angiogenesis. These results underscored the potential of the GG-g-(PDMAEA-co-PAMPS) hydrogel as an effective solution for chronic wound management.

Bacterial and helminthic evolution has resulted in a shocking development of antibiotic and anthelminthic resistance. Medicinal plants provide an encouraging source to develop new antibacterial and anthelmintic agents. Rubus ellipticus has been employed traditionally for medicinal purposes, yet its ability against microbial resistance remains to be unraveled. The present work assessed the antioxidant, antibacterial, and anthelmintic activities of leaf extracts of R. ellipticus. Antioxidant activity was measured using DPPH, antibacterial activity was done with the serial dilution technique, and anthelmintic activity was performed by using Indian earthworms. Molecular docking experiments were also carried out by utilizing Maestro 12.7 (Schrodinger) in order to screen the interaction between phytoconstituents with bacterial as well as helminthic targets. The methanol extract exhibited the highest antioxidant activity (

The colonization of fungal hyphae and spores by bacteria represents a widespread phenomenon with significant ecological and biotechnological implications across all surveyed fungal phyla. First thought to be restricted to Mollicutes and Burkholderiaceae-Related Endobacteria, these endofungal associations exhibit remarkable diversity, from simple uniform populations to complex communities, contradicting earlier assumptions of uniform populations. Acquisition dynamics demonstrate both ancient co-evolutionary relationships and recent horizontal transfer events, with environmental factors driving strain-level variation in symbiont presence even within the same fungal species. Fungi can harbor either uniform or diverse bacterial communities, sometimes within specialized structures, and exhibit varying degrees of dependence on their symbionts. These interactions can be mutualistic, commensal, or parasitic, influencing fungal physiology, metabolism, and ecological function. Yet the underlying mechanisms have been thoroughly characterized in only a few model systems in which endofungal bacteria have been shown to affect nutrient acquisition, stress tolerance, secondary metabolite production, and even fungal pathogenicity. In return, fungi offer a protective niche and promote dispersion. These concepts collectively illustrate the evolutionary flexibility and ecological importance of fungi-bacteria partnerships across terrestrial ecosystems. This review synthesizes emerging paradigms in endofungal bacteria research, integrating recent discoveries that challenge traditional assumptions about these symbioses. We examine host specificity patterns, acquisition mechanisms, and functional impacts while identifying critical knowledge gaps requiring investigation. A deeper understanding of these associations is essential to establish standardized frameworks for their applications in agriculture, medicine, and environmental sustainability.

Brassica juncea (Indian mustard) is a vital oil seed crop in India and is grown worldwide for oil and food requirements. Alternaria brassicae is a major threat to B. juncea quality and productivity causing worldwide crop loss. In this work, it was observed that exogenous melatonin (MT) at 200 mu M concentration was proven to be well effective against A. brassicae, which reduced disease consequences up to 75 %, attributed to significant reduction of necrotic lesions, delayed onset infection, and improved plant defence by enhancing chlorophyll and proline content, maintaining water retention and reducing oxidative stress. Additionally, biochemical assays, such as reduced oxidative damage, as evidenced by diminished levels of malondialdehyde (MDA), hydrogen peroxide (H2O2) and enhanced antioxidant enzyme activities, clearly showed the antifungal effect of MT on Alternaria. This study represents a new insight into the potential role as sustainable antifungal compound. Furthermore, HRMS metabolite profiling of A. brassicae cultured with MT revealed altered fungal metabolite profiles, including increased production of known antifungal compounds, suggesting that MT disrupts fungal metabolism and reduces its pathogenicity. In contrast, a decrease in the levels of antioxidant and indole derivatives in fungal extracts indicated weaker fungal defenses. The major aim of the study to provide evidence for the curative potential of MT as an effective antifungal molecule against A. brassicae in B. juncea that would be beneficial to agricultural improvement and an eco-friendly alternative to chemical fungicides.

Rice is a major dietary element for about two billion people worldwide and it faces numerous biotic and abiotic stress for its cultivation. Rice blast disease caused by Magnaporthe oryzae reduce up to 30% rice yield. Overuse of synthetic chemicals raises concerns about health and environment; so, there is an urgent need to explore innovative sustainable strategies for crop productivity. The main aim of this study is to explore the impact of bacterial volatiles (BVCs) on seedling growth and defense mechanisms of rice under in-vitro condition. On the basis of plant growth promoting properties, six bacterial strains were selected out of ninety-one isolated strains for this study; Pantoea dispersa BHUJPVR01, Enterobacter cloacae BHUJPVR02, Enterobacter sp. BHUJPVR12, Priestia aryabhattai BHUJPVR13, Pseudomonas sp. BHUJPVWRO5 and Staphylococcus sp. BHUJPVWLE7. Through the emission of bacterial volatiles compounds (BVCs), Enterobacter sp., P. dispersa and P. aryabhattai significantly reduces the growth of rice blast fungus Magnaporthe oryzae by 69.20%, 66.15% and 62.31% respectively. Treatment of rice seedlings with BVCs exhibited significant enhancement in defence enzyme levels, including guaiacol peroxidase, polyphenol oxidase, total polyphenols, and total flavonoids by a maximum of upto 24%, 48%, 116% and 80%, respectively. Furthermore, BVCs effectively promote shoot height, root height, and root counts of rice. All BVCs treated plant showed a significant increase in shoot height. P. dispersa treated plants showed the highest increase of 60% shoot and 110% root length, respectively. Root counts increased upto 30% in plants treated with E. cloacae and Staphylococcus sp. The BVCs can be used as a sustainable approach for enhancing plant growth attributes, productivity and defence mechanism of rice plant under biotic and abiotic stresses.

This study aims to evaluate the in vitro as well as in vivo antifungal activities of different phytohormones (PHs) against the hemibiotroph fungus, Fusarium oxysporum using black gram [Vigna mungo (L.) Hepper] as a model system. The potential antifungal activities were tested using PHs viz. salicylic acid (SA), methyl jasmonate (MeJA), melatonin (MT), brassinolide (BL), indole-3-acetic acid (IAA), gibberellic acid3 (GA3), ethephon (ET), and abscisic acid (ABA), by determining the minimum inhibitory concentration (MIC) and minimum effective concentration (MEC) end point in a microtiter plate-based assay. The results suggested significant antifungal activity for all the tested PHs, wherein SA and MeJA showed potency even at the lowest concentration tested, with corresponding MIC90 values of 0.312 mM and 0.625 mM, respectively. Likewise, a similar MEC profile was also observed for both SA and MeJA, with a corresponding value of 0.078 mM and 0.312 mM, respectively. The microtiter results were corroborated using spore germination and mycelial susceptibility assays. The in vivo antifungal efficacy of PHs was assessed by recording the germination characteristics in SA and MeJA-primed V. mungo seeds that were already exposed to F. oxysporum spores. The PHs-primed seeds displayed a characteristic longer seedling length and higher seed vigor index (SVI), in concomitant with relative enhanced ROS-scavenging activity. The priming of V. mungo seeds with SA and MeJA seems to induce a defense mechanism against F. oxysporum infection, which also improved its germination characteristics.

CONTEXT: Escherichia coli is one of the most important causes of urinary tract infections (UTIs). Increased antibiotic resistance may limit the therapeutic options for the treatment of E. coli infections. Fosfomycin an orally dispensed antibiotic has shown promising in vitro activity against multidrug-resistant (MDR) urinary E. coli pathogen; however, current resistance data from India are scarce.AIM: The aim of this study is to evaluate the in vitro Fosfomycin activity against uropathogenic MDR E. coli.MATERIALS AND METHODS: A total of 150 previously confirmed MDR E. coli urinary isolates were included in this study. Susceptibility testing and result interpretation of isolates to Fosfomycin was performed by the disc diffusion method as per the Clinical and Laboratory Standards Institute M100-S25 recommendations.RESULTS: Fosfomycin appears to exhibit excellent in vitro activity against the MDR E. coli urinary isolates. The susceptibility for Nitrofurantoin was fair, whereas for Ampicillin, Ofloxacin, Norfloxacin, Cefazoline and Trimethoprim/Sulphamethoxazole was found poor.CONCLUSION: In view of the high in vitro susceptibility to Fosfomycin in this population and the lack of cross-resistance between Fosfomycin and other agents, Fosfomycin may be considered a useful reserve drug in the treatment of uncomplicated UTIs caused by MDR E. coli.

Finger millet (Eleusine coracana L.) is a prevalent grain crop in the dry parts of Asia and Africa. It prolongs to be a staple food and is known locally as “Ragi” in southern states like Karnataka, Andhra Pradesh, and Telangana, especially in Karnataka. It serves as fodder as well as grain. Its grain is the richest source of calcium and it is utilized in a wide range of food products, including cakes, puddings, sweets, and other baked products. There are many homemade products prepared with finger millet and some of the well-known products are ragi roti, ragi dosa, ragi balls, ragi porridge, ragi upma, ragi cakes, and ragi biscuits. In addition, it is used to manufacture beer and liquor (known as arake or areki in Ethiopia), and also its different products are fed to animals. Due to its high fiber, mineral, vitamin, macro and micro-nutrient, phytochemicals contents, and its endowing ability to fight off chronic diseases. A cheap, satisfying, and healthful diet can be made by including finger millet in a regular diet.

Prodigiosin finds tremendous application in the food industry owing to its physicochemical and non-toxic attributes. In the present investigation, microbial production of food color (prodigiosin) was done by Serratia marcescens NCIM 5246 under solid-state fermentation (SSF). For economical prodigiosin yield, initial substrate screening was conducted using agro-waste such as rice bran, wheat bran, orange peel powder, green gram husk and cane molasses. A maximum pigment yield of 1059.67 ± 1.53 µg/L was observed on wheat bran media. Further, Taguchi Design of the experiment (DOE) using Qualitek-4 software with bigger is better as quality attributes was done for process optimization. The Taguchi (DOE) comprised five independent variables, i.e., pH, temperature, substrate concentration, trace metal concentration, and inoculum size at two levels. The software deduced independent variables’ individual and interactive effects on product yield by performing an L8 orthogonal array (OA). The output variable (prodigiosin yield) was improved based on the S/N ratio. Under optimized cultural conditions maximum prodigiosin yield of 1320.2 ± 20 µg/L was obtained. The expected prodigiosin yield of 1386.60 µg/L showed a 95

Background:The coronavirus disease 2019 (COVID-19) pandemic had highlighted the urgent need for effective preventive measures alongside conventional therapies. Ayurveda, particularly herbal fumigation (dhoopa), may offer potential complementary interventions. The Air Vaidya Herbal Dhoopa (AVHD) stick, based on traditional formulations, is evaluated for its safety and efficacy against Severe Acute Respiratory Syndrome Coronavirus 2 (SARS-CoV-2).Objectives:To assess the safety, compositional profile, and clinical efficacy of AVHD fumigation in preventing and reducing the symptoms of COVID-19.Materials and Methods:(1) Composition analysis: The AVHD stick's elemental and morphological composition was analyzed using Transmission Electron Microscopy and Scanning Electron Microscopy with Energy Dispersive Spectroscopy. (2) Clinical study: A Phase II randomized controlled trial (RCT) involving 150 subjects in the intervention group and 100 subjects in the control group was conducted, monitoring the prevalence of COVID-19-like symptoms. (3) Safety assessment: In vivo toxicity testing was performed on rodents to evaluate the safety of AVHD fumes.Results:(1) Compositional findings: Elemental analysis of the AVHD ash showed a complex mixture of elements, while the stick combined multiple medicinal plants known for antiviral and anti-inflammatory effects. (2) Clinical efficacy: The intervention group (AVHD fumigation) reported significantly lower rates of COVID-19-like symptoms compared to controls. (3) Specific symptoms reduced: Fever, cough, cold, anosmia, and loss of taste were all significantly less frequent in the intervention group. (4) Safety: Rodent toxicity studies confirmed the absence of significant adverse effects with AVHD inhalation.Conclusions:AVHD fumigation is safe and demonstrates significant efficacy in reducing the incidence of COVID-19-like symptoms, supporting its potential as complementary preventive approach against SARS-CoV-2.